气体染色学自动序列衍生 - - [轨道] 基于质谱的人类血液样本的基质代谢物分析
Akrem Jbebli1, Kateřina Coufalíková1,2, Moira Zanaboni3
1RECETOX, Faculty of Science, Masaryk University, Kotlarska 2, Brno, Czech Republic.
Bio-protocol
|March 14, 2025
概括
自动衍生简化了气色谱 (GC) 对人类血液样本的分析. 这种机器人液体处理方法提高了效率,并减少了常规代谢学研究的变化.
科学领域:
- 代谢学 代谢学 代谢学
- 分析化学 分析化学
- 生物化学 生物化学
背景情况:
- 小分子往往需要衍生用于气色谱 (GC) 分离.
- 传统的批量衍生是耗时的,并引入样本变化.
- 自动化液体处理可以提高效率,减少样品制备的变化.
研究的目的:
- 描述人类血液样本的完全自动化,两阶段衍生化协议.
- 将这种自动化协议与GC-Orbitrap质谱仪用于代谢学进行集成.
- 在常规代谢分析中提高吞吐量和可重现性.
主要方法:
- 一个机器人液体处理系统被用于连续的,自动化的衍生.
- 该协议涉及直接的甲氧化,其次是代谢产物的化.
- 样品使用GC-Orbitrap质谱分析.
主要成果:
- 实现了31分钟的样本对样本运行时间,用于高吞吐量分析.
- 在人类血液基因表中成功识别了70多种代谢物.
- 在常规测量代谢物中,经证明的定量可重复性变化系数 (CVs) 低于30%.
结论:
- 描述的自动衍生协议显著提高了效率,并减少了血液代谢学中的变异性.
- 这种方法使70多种代谢物的常规,高通量分析成为可能.
- 该协议适用于需要强大的代谢概况的临床和研究应用.
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